A5064236669- Protein purification and stability
- Monoclonal and Polyclonal Antibodies Research
- Viral Infectious Diseases and Gene Expression in Insects
- Analytical Chemistry and Chromatography
- Mass Spectrometry Techniques and Applications
- Enzyme Catalysis and Immobilization
Amgen (United States)
2020-2021
Recently, cation exchange chromatography (CEX) using aqueous volatile buffers was directly coupled with mass spectrometry (MS) and applied for intact analysis of therapeutic proteins antibodies. In our study, chemical modifications responsible charge variants were identified by CEX-UV-MS a monoclonal antibody (mAb), bispecific antibody, an Fc-fusion protein. We also report post-CEX column addition organic solvent acid followed mixing at elevated temperatures, which unfolded proteins,...
Therapeutic proteins including antibodies and Fc-fusion undergo a large number of chemical modifications during cell culture, purification, storage in human circulation. They are also exposed to harsh conditions stress studies, elevated temperature, extremes pH, forced oxidation, physiological UV light assess the possible degradation pathways suitability methods for detecting them. Some these located on residues binding regions, leading loss potency classified as critical quality attributes....
Chemical modifications (attributes) in the binding regions of stressed therapeutic proteins may affect to target and efficacy proteins. The method presented here describes criticality assessment antibody by size-exclusion chromatography (SEC) competitive between a its target, human epidermal growth factor receptor-2 (HER2), followed SEC fractionation peptide mapping characterization bound unbound antibodies. When were mixed at stoichiometric molar ratio 1:2, only antibody-receptor complex...
Intact protein mass spectrometry (MS) coupled with liquid chromatography was applied to characterize the pharmacokinetics and stability profiles of therapeutic proteins. However, limitations from chromatography, including throughput carryover, result in challenges handling large sample numbers. Here, we combined intact MS multiple front-end separations, affinity capture, SampleStream, high-field asymmetric waveform ion mobility (FAIMS), perform high-throughput specific measurements a...
Sodium dodecyl sulfate (SDS) plays a pivotal role in protein denaturation, tissue extraction, and mass-based electrophoretic separations. However, even modest concentrations of SDS can cause column overpressure, retention time shifts, ionization signal suppression during liquid chromatography-tandem mass spectrometry (LC-MS/MS) analyses. Thus, removal is critical step for LC-MS/MS analysis digests containing SDS. This study describes an inexpensive high-throughput method to remove from using...